Each antibody in the kit detects endogenous levels of its intended target.

使用抗原

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu498 of human NF-κB p65/RelA, Leu65 of human c-Rel, Ile415 of mouse NF-κB1 p105/p50, and residues near the amino terminus of human NF-κB2 p100/p52, a recombinant protein specific to the carboxy terminus of human RelB, and a synthetic phosphopeptide corresponding to residues surrounding Ser536 of human NF-κB p65/RelA.

Flow Cytometry

Western Blotting

Western blot analysis of extracts from various cell lines and tissues using c-Rel (D4Y6M) Rabbit mAb.

Western Blotting

Western blot analysis of extracts from various cell lines and rat spleen using NF-κB1 p105/p50 (D4P4D) Rabbit mAb.

Western Blotting

Western blot analysis of extracts from various cell lines using RelB (D7D7W) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

Western Blotting

Western blot anlaysis of extracts from various cell lines using NF-κB2 p100/p52 (D7A9K) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HDLM-2 cells and either RelB (D7D7W) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IAP2 promoter primers, SimpleChIP® Human IκBα Promoter Primers #5552, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

バックグラウンド

Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).

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XP is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.